Bis quaternary oximes



United States Patent 1 Claim. (Cl. 167-65) (Granted under Title 35, US.Code (1952.), see. 266) This invention described herein may bemanufactured and used 'by or for the Government of the United States ofAmerica for governmental purposes without the payment to us of anyroyalty thereon.

This is a division of application Serial No. 809,578, filed April 28,1959, now Patent No. 3,077,476, granted March 12, 1963.

This invention is directed to certain diquaternary pyri dinium halideoximes which are useful as chemothera peutic and prophylactic agents formammals poisoned by anticholinestorases, particularly the nerve gasknown as GB or sarin, i.e., isopropyl methylphosphonofluoridate.

The invention relates to l,1'-polymethylene bis (4-' formylpyridinum)halide dioximes wherein the polymeth-. ylene group contains from 2 to 6carbon atoms. These. compounds have the structural formula dioximes ofthe formula or: (N\ a C H=N O H (II) The invention also relates to1,l'-(2-butenylene) bis '(4-formylpyridinium) halide dioximes of theformula oH:NoH OH=NOH (III) The invention further relates to1,1'-polymethylene bis (3-formylpyridinium) halide dioximes of theformula CH=NOH CH=NOH (Iva) wherein R is a polymethylene groupcontaining from 2 to 6 carbon atoms.

Another class of compounds included are the 1,1'-(p- 3,093,540 PatentedJune 11, 1963 phenylenedimethylene) bis (3-formylpyridinium) halidedioximes of the formula OH=NOH (1%) This invention further relates tounsymmetrical bisquaternary 4-formylpyridini-um halide monoximes of theformula OH=NOH (V) wherein R" represents either three lower alkyl groupsor the hydrocarbon portion of the pyridine ring.

This invention also relates to the 1,l'-trimethylene bis(3-amidooximopyridinium) halides The invention also relates to thecompounds I CH=NOH (VIII) While all these compounds are useful invarying degree for the purposes set out above, the different groupsexhibit quite striking difierences in efiecti-veness. In all the aboveformulas X- is chloride, bromide or iodide, which appear to beequivalents as to physiological action, except for their eflfect onsolubility.

Wilson et al., in Patent No. 2,816,113, granted December 10, 1957,disclose a group of compounds which are effective as antidotes fortreatment of mammals poisoned by compounds of high anticholinesteraseactivity, such as the nerve gases diisopropylphosphorofluoridate (DFP),isopropyl methylphosphonofluoridate (GB), and O-ethyl, N,N dimethylphosphoroamidocyanidate (GA), as well as other related organicphosphorus compounds, including many insecticides. The compound of theWilson et al. group which has received by far the most attention is2-formyl-l-methyl pyridinium iodide oxime, commonly known as Z-pyridinealdoxime methiodide or 2-PAM. This compound is outstanding in itsability to reactivate, in vitro, acetyl cholinesterase which has beeninhibited by, for example, GB. Thus, even as compared to the veryclosely related 4-PAM, disclosed in Example II of the Wilson patent,2-PAM shows much greater activity. With isopropyl methylphosphonylatedacetylcholinesterase the rate constant at pH 7.4 and 25 C. in thepresence of 10' M acetylcholine is 2X10 per mole per minutefor 2-PAM and1.4X 10 per mole per minute .for 4-PAM. The Wilson patent shows a highrate of survival in mice which had been poisoned with paraoxon and thentreated with 2-PAM.

3 acetyl choline. Such a compound is atropine and it is at present therecommended remedy. Recently, Z-PAM has been reported to enhanceconsiderably the activity of atropine in the chemotherapeusis ofpoisoning due to or- To minimize absorption effects both the GB andTMB-4 were ordinarily given intravenously. However, in the therapeutictests on rabbits, dogs and cats, the GB was administered subcutaneously,since death from 20 LD ganophosphorus compounds, 5 intravenous dose ofGB occurs so quickly that it is virtu- The compounds of Formula I abovein which R conally impossible to give timely administration of theantitains from 2 to 6 carbon atoms are appreciably more dote. elfectivethan 2-PAM as reactivators of GB-inhibited Atropine, when administered,was included in the folacetylcholinesterase and also in enhancing theactivity of lowing amounts. atropine in both therapy and propylaxis.When X is 10 gbromide, the variation of the rate constant for the invitro R 4 reactivation of GB inhibited eel acetylcholinesterase at Raits 2 pH 7.4 and 25 C. was found to be as follows: D gs and Cats TABLE IThe prophylactic doses were given within two minutes prior to theinjection of the GB, the therapeutic doses so soon as poisoning symptomswere visible. R Rate constant Table 3 shows the results.

(l/moles/mlnutcs) 7 TABLE 3 (ohm 7x10 A. PROPHYLACTIC (CEm 6x10 (CI-I2);6x10 Grim 1x10 z-PAM TMB-4 (Orrin 6 10 A m Survival ratio An Survivalratio When administered in combination with atropine to n ms Dose lmalsDose, animals poisoned with GB the order of effectiveness was 33somewhat different. Under these conditions the comat r0- pine atropinepound in which R=(CI-I i.e., l,l-trimcthylene bis pme pine(4-formylpyridinium) bromide dioXime also known as Mi M1 12 010 -t wasmost yq- In a Challenged with a M i 3:: 40 0/6 M3313: 25 6 6 2:33:31: 2LD dose of GB adminlstered intravenously, all of a a t group of sixanimals survived if the atropine-TMB-4 com- $12 bination wasadministered intravenously immediately after poisoning. Theatropine-Z-PAM combination saved only M two of the group of animals. Onthe other hand, with dogs which were given a 20 LDso dose of GB subcuta-Miceuu 40 neously the survival ratios were the same (4/5) tor the twoRabbits- 5 treatments, which were given intravenously when syrnp- Gama40 toms appeared. However, the recovery time was much 40 shorter for thesurviving animals which received the TMB-4, i.e., 2 hours, as against 24hours for those re- The recovery periods, i.e., time for disappearanceof ceiving the Z-PAM. symptoms of poisoning, among survivors in theabove A summary of the reactivation rates and survival ratios tests,with atropine, were as follows. for these compounds when administeredtherapeutically TABLE 4 to rats together with atropine is as follows:

TABLE 2 Animals 2-PAM TMBA R Reactivation survival ProphylacticTherapeutic Prophylactic Therapeutic rate constant ratio (GB) ii ifms310 st-an i hi Egg? O ts. 5 155: 5hr 5 hr 24h OHZfi: 6x105 Dons 24hr24hr 14hr 3hr. (c1195-. 1x10 6/6 (OHQW- 6x103 3/4 The compounds ofFormula I in which R contains from 7 to 10 carbon atoms are lesseffective than those of our These Compounds constitute 0111' PresentlyPreferred preferred group. For these compounds the reactivation P- rateconstant and the survival ratio for rats (measured as our Compounds y bep y p p y y, i given above) were as follows, X- being bromide. injectedbefore exposure to the anticholinesterase agent,

e.g., GB, or therapeutically, i.e., injected subsequent to TABLE 5exposure.

The following series of experiments compares the ef- R g t l gfectiveness of our presently preferred compound, TMB-4, cons an ta 1Mwith Z-PAM applied to various animals by these two (CHm 2x103 0/4methods. The animals were poisoned by injections of GB(8H,)EIIIIIIIIIIIIIIIIIIII: 1.2 10 0/4 as follow iofiiiio'fll 1,4 103 8%Mice 0.173 mg./kg. (LDso).

Rats 0.126 mg./ kg. (2 LD While these compounds were ineffective in vivoagainst Rabbits: GB, they were, together with 2-PAM, very effectiveIntravenous 0.340 mg./kg. (20 LD against certain otheranticholinesterases, particularly that Subcutaneous 0.900 rug/kg. (20LD50)- designated as VX by the US. Army Chemical Corps. All

Dogs and cats: these compounds caused survival of all animals (survivalIntravenous 0.440 rug/kg. (20 LD rates of 4/4 and 6/6) when administeredtherapeutically Subcutaneous 0.900 rug/kg. (20 LD to rats challenged by2 LD doses of VX.

The compounds of Formula II exhibited properties intermediate those ofthe two subgroups of Formula I. When X- was chloride the compound hadthe following properties. (In this and all following tables the survivalratios are those for rats challenged by 2 LB doses of 61131 or (VX) andthe oxime was employed therapeutica y.)

Compounds of Formula III showed reactivation rates very close to thoseof our preferred group. Thus when X- in Formula III is bromide thereactivation rate constant was 8X10 as compared to the value for the R:(CH member of our preferred group of 6x10. For the unsaturated member(Formula III) the survival ratio for rats challenged by GB was only l/ 4as compared to 6/6 for the saturated analogue (Formula I). Both gavecomplete survival (ratios of 4/4 and 6/ 6) for animals challenged by VX,however.

Compounds of Formula IVa showed anomalous properties.

They gave reactivation rates which were low, but survival ratios whichwere high as compared to Z-PAM, as shown by Table 6, X- being bromide.

TABLE 6 R Reactivation Survival rate constant ratio (GB) (OHm 3. 5x10"414 (0H,); 4. 2X10 3/4 Reactivation Survival REN rate constant ratio(GB) (OzHshN 1. 2X10 4/4 The compounds of Formulas VI, VII and VIII,while being of different structure are alike in exhibiting reactivationrate constants which are very lowas compared to Z-PAM but giving highsurvival ratios as shown by Table 8, X- being bromide in each case.

TABLE 8 Formula No. Reactivation Survival rate ratio (GB) 69 4/4Negligible 4/4 67 4/4 Preparation 0 Compounds 4-pyridinecarboxaldehydeoxime was prepared by warming on a steam bath a neutralized aqueoussolution 6 of 4-pyridinecarboxaldehyde and hydroxylamine hydrochloride.The oxime had a melting point of 130130.5 C. The 2- and 3-oximes wereproduced by similar methods.

The quaternization to produce dioximes was carried out by reacting theproper oxime with a 1, n dihaloalkane, (CH ,X employing a 3:1 molarratio of oxime to halide. The unsymmetrical quaternary monoximes wereobtained be reacting the pyridine oxime with the appropriateomega-halopropyl quaternary salt in a 1.5:1 molar ratio. Two procedureswere utilized.

Procedure A: A mixture of the pyridine oxime and halide was dissolved insuflicient ethanol and refluxed for the period of time specified inTable 9.

Procedure B: A mixture of the oxime and halide was dissolved in about100 ml. of ethanol and heated in a 200 ml. capped pressure bottle(carbonated beverage type) for the length of time specified. Thereaction mixtures were cooled to room temperature and the productremoved by filtration. In several instances it was necessary to addabsolute ether to effect complete precipitation. The products wererecrystallized from ether. This procedure was usually employed becauseof its simplicity.

Table 9 gives the procedure, yields and melting points forrepresentative compounds.

TABLE 9 Melting (m) Yield or decompercent position (d) point, C.

Substituents Condi- N o. tions 208-211 in. 226-231 In. 248-251 m.280-231 d. 223-226 (1.

gases FFF QOONO Further details regarding the preparation and propertiesof certain of our compounds are given in the following publications; byus and our associates:

Pyridine Aldoxirnes by Edward J. Poziomek, Brennie E. Hackley, Jr. andGeorge M. Steinberg, Journal of Organic Chemistry, vol. 23, pp. 714-717(May 1958); and Chemotherapeutic Efiectiveness of Trimethylene Bis(4*Formyl Pyridinium Bromide) Dioxime in Anticholinesterase Poisoning,by Edmund Bay, S. Kropp and L. F. Yates, Proceedings of the Society forExperimental Biology and Medicine, vol. 98, pages 107-109 (May 1958).These articles are to be considered incorporated by reference in thisspecification.

While we have shown a number of specific examples of 7 compounds andtheir use; it will be obvious that various References Cited in the fileof this patent Poziomek: Am. Chem. Soc. Abst, 132nd Meet, 1957.

